侯兆伟,刘洋,金锐,李蔚,窦绪谋,任国领,张琨
HOU Zhaowei,LIU Yang,JIN Rui,LI Wei,DOU Xumou,REN Guoling,ZHANG Kun

• 1:国家能源陆相砂岩老油田持续开采研发中心
• 2:中国石油大庆油田有限责任公司勘探开发研究院
• 3:大庆师范学院生物工程学院

摘要(Abstract)：

在微生物降解原油产甲烷过程中，如何有效地提高微生物产甲烷能力是目前研究的技术难点之一。为了能够在短时间内利用微生物产生更多的甲烷气，提高产甲烷效率，利用污水处理厂活性污泥具有产氢和产甲烷的特性，筛选出具有稳定产氢能力的产氢菌，其菌液每100 mL产氢量可达到0.8 mmol左右；再利用产氢菌培养后产H2和CO_2以及发酵液中挥发性脂肪酸积累的特性，为产甲烷菌提供相对充足的代谢营养底物来提高产甲烷效率；通过产氢、产甲烷两阶段培养的方式，在20 d内产甲烷能力即可得到显著提升，所产气体中甲烷体积分数可达79%以上，与传统非结合方式产甲烷气体系相比，产甲烷速率提高了10倍，达到0.087 5 mmol/d，证明了应用该方法提高产甲烷效率的有效性。研究成果为进一步提高微生物降解原油产甲烷的转化效率提供了技术方向，同时也为CO_2注入产甲烷等技术的研究提供了借鉴。
In the process of methane production by microbial degraded crude oil, how to effectively improve methane production capacity of microbial is one of the technical challenges in current research. In order to use microbial to produce more methane gas in a short time and improve methane production efficiency, hydrogen producing bacteria with stable hydrogen production capacity are selected based on the characteristics of hydrogen production and methane production of active sludge in produced water treatment plant, with hydrogen production reaching about 0.8 mmol per 100 mL bacteria fluid. Then with the help of characteristics of H2 and CO_2 production by hydrogen producing bacteria after cultivation and volatile fatty acid accumulation in fermentation liquid, methane producing bacteria are provided with relatively sufficient metabolic nutrient substrates to improve methane production efficiency.By means of 2 stages cultivation of hydrogen production and methane production, methane production capacity is significantly improved in a short time during 20 d, with methane volume fraction of 79%in produced gas. Compared to traditional non-combination mode methane gas production system, methane production rate is increased by 10times to 0.087 5 mmol/d, proving the effectiveness of this method to improve methane production efficiency. The research results provide technical direction for further improving conversion efficiency of microbial degraded crude oil to produce methane, and provide reference for research on technologies such as CO_2 injection to produce methane.

关键词(KeyWords)： 活性污泥;产氢菌;产甲烷菌;氢营养型;甲烷转化率
active sludge;hydrogen producing bacteria;methane producing bacteria;hydrogen nutrition type;methane conversion rate

Abstract:

Keywords:

基金项目(Foundation): 中国石油天然气股份有限公司重大科技专项“三次采油提高采收率关键技术研究”（2021DJ1605）;; 大庆油田有限责任公司科技项目“微生物降解原油成甲烷气高转化率技术研究”（dqp-2021-sccy-ky-007）

作者(Author): 侯兆伟,刘洋,金锐,李蔚,窦绪谋,任国领,张琨
HOU Zhaowei,LIU Yang,JIN Rui,LI Wei,DOU Xumou,REN Guoling,ZHANG Kun

DOI: 10.19597/j.issn.1000-3754.202303016

参考文献(References)：

• [1]王凤兰，沙宗伦，罗庆，等.大庆油田特高含水期开发技术的进步与展望[J].大庆石油地质与开发，2019,38(5):51-58.WANG Fenglan,SHA Zonglun,LUO Qing,et al. Progress and prospects of the developing techniques in ultra-high water cut period of Daqing Oilfield[J].Petroleum Geology&Oilfield Development in Daqing,2019,38(5):51-58.
• [2]黄海平，LARTER Steve.对残余油进行生物气化以延长油田开发寿命[J].石油实验地质，2010,32(5):496-503.HUANG Haiping, LARTER S. Microbial conversion of residual oil to methane extended period of petroleum production[J].Petroleum Geology&Experiment,2010,32(5):496-503.
• [3]胡恒宇，顾贵洲，张强，等.残余油微生物气化产甲烷菌群的研究进展[J].化学与生物工程，2014,31(4):9-13.HU Hengyu, GU Guizhou, ZHANG Qiang, et al. Research progress of bacterial community for residual oil microbial gasification into methane[J]. Chemistry&Bioengineering,2014,31(4):9-13.
• [4]王立影，Mbadinga Serge Maurice，李辉，等.石油烃的厌氧生物降解对油藏残余油气化开采的启示[J].微生物学通报，2010,37(1):96-102.WANG Liying,MBADINGA S M,LI Hui,et al. Anaerobic biodegradation of petroleum hydrocarbons and enlightenment of the prospects for gasification of residual oil[J]. Microbiology China,2010,37(1):96-102.
• [5]汪卫东，王静，耿雪丽，等.储层残余油生物气化技术现状与展望[J].石油地质与工程，2012,26(1):78-81.WANG Weidong,WANG Jing,GENG Xueli, et al. Present situation and prospect of bio-gasification technology for residual oil in reservoir[J].Petroleum Geology and Engineering,2012,26(1):78-81.
• [6]冷欢，杨清，黄钢锋，等.氢营养型产甲烷代谢途径研究进展[J].微生物学报，2020,60(10):2136-2160.LENG Huan, YANG Qing, HUANG Gangfeng, et al. Recent advances in hydrogenotrophic methanogenesis[J]. Acta Microbiologica Sinica,2020,60(10):2136-2160.
• [7] SHEN L,ZHAO Q,WU X,et al. Interspecies electron transfer in syntrophic methanogenic consortia:From cultures to bioreactors[J]. Renewable and Sustainable Energy Reviews, 2016,54:1358-1367.
• [8]张国华，张志红，黄江丽.餐厨垃圾厌氧发酵连续产氢产甲烷的试验研究[J].中国沼气，2016,34(4):8-12.ZHANG Guohua, ZHANG Zhihong, HUANG Jiangli. Experiment on continuous hydrogen and methane production in food waste anaerobic fermentation[J]. China Biogas, 2016, 34(4):8-12.
• [9]刘芳，孙彩玉，边喜龙.甘蔗加工废水两相厌氧发酵产氢产甲烷性能[J].哈尔滨商业大学学报（自然科学版），2018,34(6):668-672.LIU Fang, SUN Caiyu, BIAN Xilong. Simultaneous hydrogen and methane coproduction by two stage anaerobic fermentation using sugarcane processing wastewater as substrate[J]. Journal of Harbin University of Commerce(Natural Sciences Edition),2018,34(6):668-672.
• [10]夏大平，陈曦，王闯.褐煤酸碱预处理-微生物气化联产H2-CH4的实验研究[J].煤炭学报，2017, 43(12):3221-3228.XIA Daping,CHEN Xi,WANG Chuang. Experimental study on the production of H2-CH4 from lignite jointly with acid-alkali pretreatment-microbial gasification[J]. Journal of China Coal Society,2017,43(12):3221-3228.
• [11]任国领，徐晶雪，张莹，等.不同激活剂对油藏内源微生物群落结构的影响[J].中国微生态学杂志，2015,27(9):1004-1008.REN Guoling,XU Jingxue,ZHANG Ying,et al. Effect of different activator systems on oil reservoir indigenous microbial community structure[J]. Chinese Journal of Microecology,2015,27(9):1004-1008.
• [12]解军，祁峰，裴海燕，等.脱氢酶活性检测方法及其在环境监测中的应用[J].中国环境监测，2006,22(5):13-18.XIE Jun, QI Feng, PEI Haiyan, et al. Determining method of dehydrogenase activity and its application in environmental monitoring[J]. Environmental Monitoring in China, 2006, 22(5):13-18.
• [13]李秀芬，胡庆昊，陈坚.不同底物条件下金属离子螯合剂对厌氧消化的影响[J].环境科学，2009,30(6):1701-1704.LI Xiufen, HU Qinghao, CHEN Jian. Effect of metal chelating agent on anaerobic digestion under different substrates[J].Environmental science,2009,30(6):1701-1704.
• [14]任南琪，王爱杰，马放.产酸发酵微生物生理生态学[M].北京：科学出版社，2005.REN Nanqi, WANG Aijie, MA Fang. Physiological ecology of acidogenic fermentation microorganisms[M]. Beijing:Science Press,2005.
• [15]昌盛，李建政，刘枫，等.不同热处理时间对污泥厌氧发酵产氢的影响[J].环境工程技术学报，2015,5(1):7-12.CHANG Sheng, LI Jianzheng, LIU Feng, et al. Effect of heat pretreatment time on seed sludge for fermentative hydrogen production[J]. Journal of Environmental Engineering Technology,2015,5(1):7-12.
• [16]张立国，艾冰凌，李建政，等.氧化还原介体强化厌氧活性污泥发酵产氢特征[J].中国环境科学，2021,41(5):2196-2202.ZHANG Liguo,AI Bingling,LI Jianzheng,et al.Improving fermentative hydrogen production of anaerobic sludge by redox mediators[J]. China Environmental Science, 2021, 41(5):2196-2202.
• [17]刘飞，方柏山.微生物产氢机理的研究进展[J].工业微生物，2007,37(3):58-62.LIU Fei,FANG Baishan. Research advance in biohydrogen mechanism[J]. Industrial Microbiology,2007,37(3):58-62.
• [18]刘洋.微生物降解原油产天然气室内研究[J].中外能源，2015,20(1):37-42.LIU Yang. Laboratory study on natural gas produced by microbial degradation of crude oil[J]. Sino-Global Energy, 2015, 20(1):37-42.
• [19]承磊，郑珍珍，王聪，等.产甲烷古菌研究进展[J].微生物学通报，2016,43(5):1143-1164.CHENG Lei, ZHENG Zhenzhen, WANG Cong, et al. Recent advances in methanogens[J]. Microbiology China, 2016, 43(5):1143-1164.
• [20]刘春艳，于建青，江小青.生物气形成过程中氢元素的来源[J].承德石油高等专科学校学报，2014,16(4):5-8.LIU Chunyan,YU Jianqing,JIANG Xiaoqing. Characteristics of hydrogen stage in bio-methane production[J]. Journal of Chengde Petroleum College,2014,16(4):5-8.